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Involvement of membrane sulfhydryls in the activation and maintenance of nutrient transport in chick embryo fibroblasts.

At 5 microgram/ml, insulin stimulates hexose, A-system amino acid, and nucleoside transport by serum-starved chick embryo fibroblasts (CEF). This stimulation, although variable, is comparable to that induced by 4% serum. The sulfhydryl oxidants diamide (1-20 micrometer). hydrogen peroxide (500 micrometer), and methylene blue (50 micrometer) mimic the effect of insulin in CEF. PCMB-S,1 a sulfhydryl-reacting compound which penetrates the membrane slowly, has a complex effect on nutrient transport in serum- and glucose-starved CEF. Hexose uptake is inhibited by 0.1-1 mM PCMB-S in a time- and concentration-dependent manner, whereas A-system amino acid transport is inhibited maximally within 10 min of incubation and approaches control rates after 60 min. A differential sensitivity of CEF transport systems is also seen in cells exposed to membrane-impermeant glutathione-maleimide I, designated GS-Mal. At 2 mM GS-Mal reduces the rate of hexose uptake 80-100% in serum- and glucose-starved CEF; in contrast A-system amino acid uptake is unaffected. D-glucose, but not -L-glucose or cytochalasin B, protects against GS-Mal inhibition. These results are consistent with the hypothesis that sulfhydryl groups are involved in nutrient transport and that those sulfhydryls associated with the hexose transport system and essential for its function are located near the exofacial surface of the membrane in CEF.

Amino Acids

Loss of the post-translational control of nutrient transport in vitro and in vivo virus-transformed chicken cells.

The removal of serum from the medium of uninfected fibroblasts decreased the rate of uptake of uridine, 2-deoxyglucose, alpha-aminoisobutyrate and thymidine. Its subsequent addition rapidly and reversibly stimulated the uptake of all the nutrients but thymidine and this response was not inhibited by treatment of the cells with cycloheximide. The cycloheximide insensitive, rapid increase in the rate of transport has been designated post-translational control. The nutrient transport systems in chick embryo fibroblasts transformed in vitro with avian sarcoma viruses and virus-induced cultured chicken tumor cells do not respond to serum removal or addition. Two possible levels for the control of nutrient transport, i.e., mitogen receptor occupancy and mitrogen-induced activation of the transport system, are presented to explain these observations.

Aminoisobutyric Acids

Insecticide and nutrient transport in water, related to agricultural land use of a stream basin in Ontario, Canada.

Transport by stream water of insecticides and nutrients in Big Creek, Norfolk County, Ontario, Canada, was examined by combining concentrations of substrates with flow data. Big Creek has its headwaters in dairy cattle country, its central basin area is mainly devoted to tobacco growing, and its lower reaches contain mixed farming, corn and vegetables, etc., before it flows into Lake Erie. Three sampling sites were chosen to represent these 3 different land uses. Generally concentrations of substrates in the water were quite uniform at the 3 sites resulting in transport of quantities in proportion to stream flow. Certain anomalies occurred and are discussed. The 2 chief insecticides found were DDT and dieldrin. Analyses for potassium, calcium and magnesium indicated that these nutrient losses into the stream were area-rather than usage-dependent. Midseason variations in loss of nitrogen and phosphorus may be the result of agricultural practices in the three areas represented in this study. The largest quantities of all nutrients lost occurred early in the season before crops were established.

Agriculture

Effect of cations on intestinal nutrient transport in two teleosts.

The uptake of sugar and amino acids was affected by the presence of cations in the filling solution in both the fishes, Ophiocephalus and Heteropneustes. Under low Na+ concentration, the rate of transport decreased while an increase in Na+ concentration brought about its corresponding increase in both the fishes. Li+ was able to substitute Na+ to some extent in the filling solution in the transport of xylose, glycine and leucine. The replacement of Na+ by Li+ was more successful in xylose transport, in contrast to the transport of glycine and leucine. On the other hand, K+ was not able to substitute Na+ in the transport process. K+ inhibited the transport of glycine but did not that of xylose and leucine.

Animals

Nutrient transport systems in dog brain.

The cerebral circulation of the dog was isolated and an extracorporeal system was used to perfuse the brain with diluted blood. Transport of blood-borne substances was investigated by measurement of arteriovenous (a-v) differences (net transport) and by the indicator dilution method (unidirectional influx). The net movement of amino acids between blood and brain was qualitatively and quantitatively different when measured by whole blood a-v differences than when measured by plasma a-v differences. This result suggests that erythrocytes are involved in the movement of amino acids between the blood and brain, a finding similar to that observed in other vascular beds (Elwyn et al. Am. J. Physiol. 222: 13338 1972). The transport of 13 different blood-borne substances was investigated at normal and elevated solute concentrations using the indicator dilution method. Saturable processes were observed for the unidirectional transport of tyrosine, tryptophan, L-dopa, free palmitic acid, adenine, thymine, adenosine, and guanosine. However, the blood-brain interface did not appear to contain a carrier for dopamine, folic acid, cyanocobalamin, guanine, hypoxanthine, and free oleic acid.

Adenine

Transport of nutrients by a thermophilic bacterium--reconstruction of vesicles from crystalline ATPase or solubilized alanine carrier.

A strain of aerobic thermophilic bacteria was selected in order to purify highly stable membrane proteins and no reconstitute proteoliposomes capable of transporting nutrients from them. These proteins responsible for the transport could be divided into (1) proteins which supply energy to the transporting system, and (2) specific nutrient carriers driven by the energy. The former included a stable ATPase (TF1) and a lipoprotein TF0) which rendered TF1 sensitive to energy transfer inhibitors. The complex of TF0 anlysis of ATP. And one of the latter reported in this paper was alanine carrier protein which was driven by proton movement. TF1 was the first crystallized ATPase in biomembranes, and was reconstituted from its five different polypeptides, two of which were necessary for ATPase activity and four of which, for proton translocation. Purification of alanine carrier and reconstitution of proteoliposomes capable of alanine accumulation were also demonstrated.

Adenosine Triphosphatases

Placental transport of nutrients.

The fetus is dependent upon the placenta for transport to it of all nutrients for energy and growth. The primary nutrients crossing the placenta are glucose, lactic acid, free amino acids, free fatty acids, and ketone bodies. Under normal circumstances, it appears that glucose is the primary energy source. In abnormal conditions there is the possibility that energy requirements may be met also by protein-amino acid oxidation and ketone body metabolism. The fetal brain may be quite adept in the use of ketone bodies. In all transport phenomena, the placenta intervenes by diverting nutrients into its own intermediary metabolism. For a more extensive consideration of these factors, several in-depth reviews are recommended. To better understand abnormalities of intrauterine growth, we need much more specific information about the mechanisms of placental transport and the intermediary metabolism of the placenta and fetus. When these are adequately in states of normal and abnormal intrauterine growth, we can then consider methods of metabolic intervention which will correct intrauterine growth failure and, hopefully, eliminate its hazards to the fetus and child.

Amino Acids

Sugar and amino acid transport in animal cells.

The molecular basis of intracellular metabolism of nutrients and its control is quite well understood in animal cells. Comparable knowledge about solute entry into cells is still lacking, as, in contrast to metabolism, no chemical reactions seem to be directly associated with the known nutrient transport. Nevertheless, translocations of sugars and amino acids across the plasma membrane are specific and controlled processes, biologically as well as chemically. Recent advances in techniques for isolation of plasma membranes have made it feasible to study transport properties of animal cells without the complications encoutered in viable cells. This approach has been applied to sugar and amino acid transport in plasma membranes of several tissues, and intact transport systems for D-glucose, D-fructose, neutral L-amino acids, and dipeptides have been demonstrated. This demonstration of intact transport systems in an in vitro setting accomplishes the first step in the direction of molecular isolation of transport systems. Furthermore, the information obtained about the transport mechanism catalyzed by some systems has settled controversies on active nutrient transport. For example, electrogenic cotransport of sodium and D-glucose or of sodium and neutral L-amino acids has been shown to form the basis for active, sodium-dependent absorption of these nutrients. A consequence of this type of mechanism is interaction between sugar and amino acid transport via the common charged cosubstrate sodium. Moreover, different types of transport systems for the same substrate have been demonstrated in the luminal and contraluminal regions of the plasma membrane of epithelial cells, which explains unidirectional transepithelial transport. The luminal membrane contains sodium-dependent, active transport systems, and the contraluminal membrane passive, facilitated diffusion systems. In vivo, the lower intracellular sodium potential would result in concentrative nutrient uptake from the lumen, but would not influence exit on the contraluminal side. Variations in the electrical components of the sodium potential, which have not been measured, may explain apparently contradicting results on active sugar and amino acid transport with various tissue preparations.

Amino Acids

Effect of Chang'an decoction on ulcerative colitis by regulating T helper 17 cells and regulatory T cellsRab27 in the p53/high mobility group box 1 pathway.

OBJECTIVE: To explore the effect of Chang'an decoction (, CAD) of ameliorating the immune imbalances in ulcerative colitis (UC) by regulating Rab27 in the P53/high mobility group box 1 pathway. METHODS: The functions and important signaling pathways of the Rab27- and UC-related genes were analyzed viathe use of microarray data from the gene expression omnibus database, gene ontology database, Kyoto encyclopedia of genes and genomes database and gene set enrichment analysis. Dextran sulfate sodium salt-induced colitis mouse model was used to verify the bioinformatics results. Colon length, body weight, and disease activity index were measured. Hematoxylin and eosin staining was applied to validate the histopathology. Tight junction proteins were detected by immunohistochemistry. The proportions of T helper 17 cells (Th17) and regulatory T cells (Treg) in mesenteric lymph nodes were measured viaflow cytometry. Proinflammatory cytokines like interleukin (IL) 17 (IL-17), IL-21 and IL-22 and anti-inflammatory cytokines like transforming growth factor β and IL-10 in the serum and colon of mice were detected by enzyme-linked immunosorbent assay and quantitative real-time polymerase chain reaction, respectively. The expression levels of high mobility group box 1 (HMGB1), P53 and phospho- P53 (P-P53) in colonic tissues were detected by immunofluorescence and Western blotting. RESULTS: Bioinformatics analysis revealed that compared with normal tissues, the expression of Rab27 was significantly increased in UC tissues. Receiver operating characteristic curve showed that Rab27 has the potential to be used as a biomarker for the diagnosis of disease activity. Enrichment analysis showed that UC and Rab27 were mainly associated with small molecule transport, nutrient metabolism, transmembrane transport and the downstream pathway of P53. According to animal experiments, the expression of Rab27 was increased in UC tissues, which aggravated the colonic pathological damage, activated the expression of HMGB1, and also leaded to the imbalance of Th17 and Treg cells. After CAD intervention, Rab27 overexpression, weight loss, colon shortening, and pathological damage were substantial reduced, the expression of tight junction proteins, zona occludens 1 and Occludin were increased. The effect of CAD at high-dose was more obvious. In addition, CAD upgraded the number of Treg cells and the production of TGF-β and IL-10, while decreasing the number of Th17 cells and the expression of inflammatory cytokines (IL-17, IL-21, and IL-22). Moreover, colon inflammation was alleviated by CAD, as indicated by the regulation of HMGB1 and P-P53 expression. CONCLUSION: The expression of Rab27, HMGB1 and P-P53 could be decreased by CAD, and the balance of Th17 and Treg cells as well as their related cytokines could be regulated by CAD.

Animals

Cyclic AMP, membrane transport and cell division. I. Effects of various chemicals on cyclic AMP levels and rate of transport of neucleosides, hypoxanthine and deoxyglucose in several lines of cultured cells.

Nutrient transport rates and cyclic AMP levels have been implicated in the regulation of cell proliferation. In the present study, however, changes in intracellular cyclic AMP level in several lines of cultured cells (normal 3T3 and SV40 and polyomavirus-transformed 3T3 cells; 3T6, C6 GLIOMA, MOUSE L, and Novikoff rat hepatoma cells) by treatment with papaverine, prostaglandine E1 or isoproterenol did not correlate with the inhibition of the uridine, hypoxanthine or deoxyglucose transport rates by these chemicals. Transport inhibitions by above chemicals or Persantin or Cytochalasin B occurred in most cell lines in the absence of any measurable change in intracellular cyclic AMP concentration. Furthermore, treatment of several cell lines with 1 mM dibutyryl cyclic AMP had no immediate effect on the transport of uridine, thymidine or deoxyglucose, although the transport capacity of the cells for uridine and thymidine, but not that for deoxyglucose, decreased progressively with time of treatment. We also observed that the uridine transport system of all cell lines derived from 3T3 cells and the hypoxanthine transport system of L cells exhibited high degrees of resistance to inhibition by the various chemicals. On the other hand, deoxyglucose transport was inhibited to about the same extent by these chemicals in all the cell lines investigated.

Animals

Toxic substances and cell membrane function.

The exposed location and functional importance of cell membranes make them particularly susceptible to the toxic effects of many chemicals. The likelihood of such effects has been appreciated for many years. However, the recent advent of new techniques has greatly increased our understanding of the complexities of membrane structure and function. These data make it quite clear that the interaction of toxic compounds with either the protein or the lipid component of cell membranes may substantially alter membrane function. This paper summarizes the current concepts of membrane structure and function and discusses the techniques currently in use to study cell membranes. Several examples are presented in which xenobiotics significantly alter membrane function. These include effects of heavy metals on passive ion permeability, impairment of osmoregulation and calcium transport by organochlorine pesticides, inhibition of the transport of neurotransmitter metabolites by phenoxyacetic acid herbicides in choroid plexus, and reduction in intestinal nutrient transport by heavy metals. Hence the study of the interactions of foreign compounds with membrane function may enhance our understanding of mechanisms both of toxicity and of basic membrane function.

2,4-Dichlorophenoxyacetic Acid

Balancing nutrient remobilization and photosynthesis: proteomic insights into the dual role of lupin cotyledons after germination.

Efficient nutrient mobilization from seed storage tissues is essential for seedling establishment, particularly in legumes such as Lupinus albus (white lupin), which thrive in nutrient-poor soils. This study investigates the role of cotyledons in nitrogen (N) and mineral remobilization after germination during their transition from storage organs to photosynthetically active tissues, including the metabolic challenges posed by the coexistence of these two functions in epigeal germination. We cultivated white lupin seedlings under nitrogen-deficient conditions, analyzing cotyledon composition and function over 28 days. Our results indicate that 60% of cotyledon-stored proteins are degraded within the first 8 days, with free amino acids transiently accumulating before being redistributed to support growth. The progressive depletion of cotyledon reserves was accompanied by structural and metabolic changes, including an increase in photosynthetic proteins. However, cotyledon photosynthetic capacity remained lower than that of true leaves, suggesting a transient role in energy metabolism. The loss of cotyledons before day 12 significantly impaired seedling development, emphasizing their critical contribution to nitrogen, phosphate, and micronutrient supply during early growth. Comparative proteomic analysis revealed dynamic shifts in nutrient transport, amino acid metabolism, and stress response pathways following cotyledon removal. These findings underscore the significance of cotyledon nutrient remobilization in legume adaptation to low-fertility soils and highlight potential targets for breeding strategies aimed at improving nutrient use efficiency. By optimizing cotyledon nutrient composition and function, future breeding efforts could enhance seedling vigor, reduce fertilizer dependency, and improve the nutritional value of lupin-based foods.

Lupinus

Plasma membrane vesiculation: a cellular response to injury.

The shedding of plasma membrane vesicles has been shown to result from exposure of monolayer cell cultures to formaldehyde and other sulfhydryl blocking agents. Incubation of cells in concentrations of these agents as low as 5 to 10 mM for intervals as brief as fifteen minutes is effective (Scott, 1976). Plasma membrane vesiculation has been shown to be an energy-dependent process that requires Ca++ and physiological temperature. Following plasma membrane vesiculation, cell monolayers appear intact by phase microscopy and show only slight evidence of cell injury by electron microscopy. In view of these observations, the question has been raised whether plasma membrane vesiculation is compatible with continued cell growth and metabolism. The experiments described in this paper were designed to answer these questions. We pulse exposed 3T3 mouse embryo cells to concentrations of formaldehyde, between 2.5 and 250 mM, for intervals 15, 30 or 60 min. Cell momolayers were then washed in a variety of different media in an attempt to reverse the effect of formaldehyde on cells. Cell monolayers were thereafter assayed for the shedding of plasma membrane vesicles and for their ability to transport 2-deoxy-D-glucose. Cells were also replated in serum-containing medium and their ability to grow was assayed over a seven day interval. The results show an inverse relationship between the shedding of plasma membrane vesicles and the ability of the cells to transport nutrients and to grow. We interpret these data to suggest that the process of plasma membrane vesiculation results from a form of cell injury which blocks cellular metabolism and growth.

Biological Transport, Active

Metabolic reprogramming and taxonomic drivers in bacterial vaginosis: A large-scale metagenomic meta-analysis.

OBJECTIVE: Bacterial vaginosis (BV) represents a profound ecological shift from a Lactobacillus-dominated microbiota to a diverse polymicrobial biofilm associated with adverse outcomes. While taxonomic signatures are well-documented, the functional mechanisms driving this transition remain obscured. This study elucidates the genomic potential for metabolic reprogramming and the putative "functional handover" underpinning the stability of the dysbiotic state. METHODS: A computational meta-analysis of 3557 vaginal microbiomes from diverse global cohorts was performed using the standardized MGnify pipeline. A high-resolution subset of 187 whole-genome shotgun (WGS) metagenomes was stratified to compare functional potential across demographic groups. Taxon-function interaction networks were constructed, utilizing a dual-filter statistical approach (p&#x202f;<&#x202f;0.05 and effect size ranking), to map the shift from homeostatic maintenance to dysbiotic metabolic potential. RESULTS: BV was characterized by a fundamental shift from "maintenance" pathways to high-turnover "growth-oriented" genomic repertoires. While ABC transporter-like domains were present in healthy communities, dysbiosis was marked by a quantitative expansion and diversification of these systems alongside P-loop NTPases. Network analysis revealed a putative "functional handover": while Gardnerella serves as the adherent structural scaffold, the metabolic burden appears to be associated with secondary anaerobes, specifically BVAB1 and Sneathia, which exhibit strong genomic correlations with nutrient transport and stress response pathways. Crucially, microbiomes from women of African ancestry (Black cohort) exhibited a distinct functional profile with genomic signatures consistent with functions previously associated with resistome expansion (e.g., tetracycline/macrolide resistance), contrasting with Asian cohorts. CONCLUSION: BV is a state of metabolic reprogramming where genomic functional dominance is transferred from Lactobacillus to a cooperative network of anaerobic opportunists. Identifying BVAB1 and Sneathia as candidate metabolic engines, supported by a Gardnerella scaffold, challenges current therapeutic paradigms and highlights the potential for precision medicine targeting specific functional drivers and resistome profiles across diverse populations.

Humans

Flow of reducing equivalents into isolated intestinal mitochondria.

A system of enzymes is required for the transport of reducing equivalents from reduced nicotinamide adenine dinucleotide (NADH) generated in the cytosol into the mitochondria by the substrate cycles. Also, the intestinal mitochondria must be capable of oxidizing the substrates of the cycles. Both substrate cycle enzymes and permeable mitochondria are necessary for the flow of pyruvate derived from glucose into the mitochondria for oxidative decarboxylation and for the efficient production of adenosine 5'-triphosphate (ATP) for the unique intestinal nutrient transport functions. Mitochondria from hamster intestinal mucosa were prepared exhibiting good respiratory control ratios. The isolated intestinal mitochondria would not oxidize NADH unless N,N,N',N'-tetramethyl-p-phenylenediamine (TMPD) was added as a carrier of reducing equivalents. The rates of oxidation of the substrates of the L-glycerol 3-phosphate and the L-malate/1-aspartate substrate cycles were measured with the mitochondria isolated from the small intestinal mucosa. The key enzymes measured in the cytosol and mitochondria from the mucosa were NAD-L-glycerol 3-phosphate dehydrogenase, Fp-L-glycerol 3-phosphate dehydrogenase, L-malate dehydrogenase and L-glutamate-oxaloacetate transaminase. In addition, the substrate cyclase were simulated in vitro by following NADH oxidation by isolated mitochondria in the presence of added cytosolic constituents.

Animals

Hologenomic insights into the molecular adaptation of deep-sea coral Bathypathes pseudoalternata.

Deep-sea coral ecosystems support biodiversity and nutrient cycling through interactions with symbionts. However, their molecular mechanisms remain unexplored. Here, hologenomic analyses of Bathypathes pseudoalternata are applied to uncover molecular adaptations underpinning host-symbiont interactions. Genomic evidence reveals that B. pseudoalternata exhibits adaptations in nutrient transport, immune response, and lysosomal digestion, reflecting its genomic adjustments for a stable symbiosis. Candidatus Nitrosopumilus bathypathes (78.43% &#xb1; 3.65%) is inferred to oxidize host-derived ammonia to synthesize amino acids and vitamins to provision the host. The presence of CRISPR-Cas and restriction-modification (R-M) systems suggests that Ca. Bathyplasma bathypathes and Ca. Thalassoplasma bathypathes (10.68% &#xb1; 2.99%) may protect the host from viral infections. Ca. Bathybacter bathypathes (8.39% &#xb1; 1.53%) is hypothesized to synthesize heme, lipoic acid, and glutathione, which serve dual functions as antioxidants and nutrients. These findings collectively provide insights into how the hologenome contributes to the survival of B. pseudoalternata in the extreme environment.

Animals

Microbial decomposer diversity and metabolic function during the decomposition of brine shrimp carcasses in a saline lake.

BACKGROUND: Decomposition of brine shrimp carcasses has a crucial role in carbon cycling of saline lakes, yet the microbial dynamics remain poorly understood. RESULTS: Here we integrated metagenomics, metatranscriptomics, culturomics, metabolomics, and microcosm experiments to investigate microbial community succession and function during brine shrimp (Artemia sp.) carcass decomposition in Barkol Lake, a hypersaline lake in China. A total of 149 metagenome-assembled genomes (MAGs) and 77 pure culture genomes were recovered across 33 phyla, with 72.12% genomes representing species-level novel lineages. Our results reveal diverse bacterial and archaeal taxa, including novel lineages from CG03, T1Sed10-126 and rare archaeal taxa (Asgardarchaeota, Thermoplasmatota, Nanoarchaeota, and Halobacteriota), involved in degradation of biomacromolecules-proteins, carbohydrates, lipids, and nucleic acids-via extracellular hydrolysis, nutrient transport, and intracellular catabolism. These taxa exhibit substrate preferences, rapidly responding to the breakdown of polysaccharides and proteins, followed by lipids and nucleic acids. Hydrolyzed oligomers are further oxidized by various microbes through fermentation, sulfate reduction, and methanogenesis via metabolic handoffs. Additionally, viral auxiliary metabolic genes (AMGs) further enhance microbial host functions, contributing to key ecological processes such as carbon cycling and stress response. A temporally structured microbial decomposer network (MDN) was observed, driving mineralization cascades from fermentation to sulfate reduction and methanogenesis. CONCLUSIONS: This study reveals microbial metabolic handoffs and virus-mediated modulation as critical mechanisms for organic matter turnover, expanding the known diversity and function of decomposers in saline ecosystems. Our findings offer new insights into biogeochemical processes in saline lakes and highlight a synergistic microbial decomposer network involving bacteria, archaea, and viruses that collectively drive nutrient cycling during brine shrimp carcass decomposition. Video Abstract.

Animals